Evaluation of three methods for high throughput extraction of DNA from challenging fish tissues

Three methods for extracting DNA were tested on otoliths, scales, fins, and gill tissue from European whitefish [Coregonus lavaratus (L.)]. The aim was to find time-efficient and affordable ways to simultaneously extract DNA suitable for conservation genetic studies from a large number of samples and different tissues. A rapid low-cost method led to 97 % success of microsatellite amplification in otoliths and 100 % in scales. High amplification success was achieved with fin (97 %) and gill (99 %) tissue using a salt lysis-based protocol. A commercial extraction kit delivered good results with all tissues. The findings are useful for conservation genetic studies using both contemporary and archived samples.     

Obstruction after Burch colposuspension: a return to retropubic urethrolysis

We compare our postoperative outcomes of a vaginal vs abdominal urethrolysis after an obstructed Burch colposuspension. We performed a retrospective review of all women who had undergone a Burch urethrolysis from 1997 through 2003. Data collected included pelvic examination, cystoscopic and urodynamic findings, surgical morbidity, and pre- and postoperative urogynecologic symptomatology. Sixteen women underwent Burch urethrolysis. Preoperative symptoms were obstructive in 6, overactive in 2, and both (obstructive/overactive) in 8. All had an acute retropubic angulation of the urethral axis as well as a retropexed urethra that prohibited sagittal rotation of a rigid cystoscope. Seventy-eight percent of the women with urodynamic information met urodynamic criteria for urethral obstruction. Average follow-up was 7 months, and of those undergoing a vaginal urethrolysis, three out of seven (43%) had resolution of their obstructive or overactive symptoms postoperatively. In contrast, seven out of nine women (78%) who underwent a retropubic urethrolysis had relief of their obstructive or overactive bladder symptoms. In our cohort study, an abdominal retropubic urethrolysis was more successful in relieving urethral obstruction than a vaginal urethrolysis.     

Overt rectal prolapse following repair of stage IV vaginal vault prolapse

Pelvic organ prolapse is an increasingly common problem as women are living longer. With the growing numbers of surgeries performed to correct this problem, further research is needed to understand the long-term success as well as possible complications of these procedures. One potential complication that needs further study is de novo rectal prolapse after repair of pelvic organ prolapse, specifically after colpocleisis. Defacography may be an important part of the preoperative workup in the patient with pelvic organ prolapse. Currently, there is a controversy as to whether internal, or occult, rectal prolapse on defacography should be repaired at the time of other pelvic reconstructive surgery. We report on a case of overt rectal prolapse after repair of Stage IV vaginal vault prolapse with a colpocleisis, levator ani plication, and a minimally invasive midurethral sling. We discuss the issues surrounding preoperative management of these patients and propose a theory explaining why prolapse in other areas of the pelvis may occur after reconstructive surgery.     

Phosphoinositide 3-kinase is involved in the induction of the human sperm acrosome reaction downstream of tyrosine phosphorylation

In somatic cells phosphoinositide 3-kinase (PI 3-kinase) is activated upon interaction with both receptor tyrosine kinases (RTK) and G- proteins resulting in the production of moieties involved in the inositol phospholipid signalling pathway. As G proteins, RTK and the inositol phospholipids have all been implicated in the human sperm acrosome reaction, experiments were carried out to determine whether PI 3-kinase was also involved in this phenomenon. Wortmannin is a selective inhibitor of PI 3-kinase and was shown to significantly inhibit the acrosome reaction induced by both mannose-bovine serum albumin (mannose-BSA) (10, 50 and 100 nM) and a polyclonal antibody raised against an extracellular region of the sperm zona receptor kinase (ZRK, at 100 nM only). Wortmannin did not inhibit the A23187- or progesterone-induced acrosome reaction. These results suggest that PI 3- kinase is involved in the human sperm acrosome reaction. The levels of tyrosine phosphorylation of sperm proteins as detected by Western blotting using antiphosphotyrosine antibodies was not affected by wortmannin in agonist (A23187 and mannose-BSA)-stimulated spermatozoa. This indicated that PI 3-kinase operates downstream of tyrosine phosphorylation in the signal transduction cascade which leads to the human sperm acrosome reaction.      

Magnetic resonance angiography

Pulse sequences that permit selective detection of moving spins in a magnetic resonance image have been developed. Experiments were performed by the authors to produce projected angiographic data without the use of contrast agents, with the intensity of each image pixel determined by the macroscopic velocity of the detected spins. With this method, suppression of nonmoving spins is essentially complete, yielding a high dynamic range in signal intensity for detected vessels. Selective detection of moving spins is not dependent on pulsatile flow. Consequently, not only arterial structures, but also venous structures can easily be visualized. High-resolution angiographic images can be obtained by combining the flow experiment with surface coil techniques.     

Pharmacokinetics of apomorphine in parkinsonian patients

Summary— Apomorphine, a dopamine agonist, has been used efficiently in parkinsonian patients to treat severe levodopa-induced on-off phenomenon. Motor improvement has been obtained both with continuous subcutaneous (SC) infusions, and multiple SC injections. So as to assist in the understanding of the clinical results, we studied the peripheral pharmacokinetics of apomorphine in 20 patients after intravenous (IV) or SC injections in the anterior abdominal wall and in the thigh at various doses, or SC infusion. Plasma apomorphine levels were measured by high-performance liquid chromatography with electrochemical detection. After an SC injection in the abdominal wall, the T_max was brief (16 ± 11 min) the drug was rapidly cleared from the plasma and had a short plasma half-life (69.7 ± 25.8 min). The AUC was similar following SC and IV injections, suggesting that apomorphine was completely absorbed from subcutaneous tissue. Inter-subject variability in drug absorption was large. We noticed a trend towards a more complete absorption following injection in the abdominal wall rather than in the thigh. In patients chronically treated by continuous SC infusion, the apparent plasma half-life was five times longer than that following SC or IV injections. These pharmacokinetic data may explain the rapid onset and brief duration of clinical effects, and the usefulness of individual titration for intermittent SC apomorphine injections, and the smoother motor response obtained with continuous SC infusions.     

Current and future techniques of neuromodulation for bladder dysfunction

Recent increase in the use of neuromodulation for the treatment of urinary urgency and frequency, urge incontinence, and nonobstructive urinary retention has coincided with an improved knowledge of micturition physiology and concurrent technologic advances in nerve stimulation. Currently, the most common technology for bladder neuromodulation involves stimulation through the S3 foramen, or sacral neuromodulation, although other techniques of neuromodulation continue to be explored. Despite many advances in neuromodulation, the exact mechanism of action remains uncertain. Additionally, which patients will respond to neuromodulation remains unclear, and although there is a standard method for testing, this procedure is invasive and expensive. As we continue to improve patient selection criteria and better understand the mechanism of action, the efficacy and patient satisfaction should continue to increase. Currently, most patients considered for implantation with a neuromodulator are those unresponsive to other conservative treatments for bladder dysfunction.